Short protocol:
1:5000 dilution in 5% low fat milk powder in PBS-T and incubate overnight at 4°C.
Dynabeads protein A kits (Invitrogen) were used for IP. Longer incubation of the antibody in the Ab binding and washing buffer of 1 h was used for optimal binding.
Review:
Worked perfectly for both immunoblotting and co-IP. The diluted antibody could be stored at -20°C after use and be reused 5-10 times for immunoblotting.
Western blot
Immunostaining
Not Rated
IP
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
On March 16, 2016Sune Skeldal, Aarhus University wrote:
5
Good antibody against His-tag for western blotting. Detected his-tagged constructs expressed in media from CHO cells with good result. I used 1:2500 dilution.
Western blot
Immunostaining
Not Rated
IP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
On November 4, 2015Camilla Gustafsen, Aarhus University wrote:
4
This antibody (1:2500) specifically recognizes 6His tagged PCSK9 secreted to the media of stably transfected HEK293 cells. No band is detected in the lane loaded with media from cells transfected with un-tagged PCSK9. However, a strong background signal is detected from the SeeBlue Plus2 pre-stained protein marker. See inserted figure
Western blot
Immunostaining
Not Rated
IP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
On June 3, 2015Camilla Gustafsen, Aarhus University wrote:
5
We have used this antibody for WB detection of His tagged PCSK9 produced in CHO cells.
Great result. No background signal
We dilute 1:2500 in 5 % milk buffer and reuse several times
Western blot
Immunostaining
Not Rated
IP
Not Rated
ELISA
Not Rated
Flow cytometry
Not Rated
Luminex
Not Rated
On February 14, 2012Simon Glerup, Aarhus wrote:
3.3
Reasonable for detecting His-tagged proteins by Western blotting.
WB dilution 1:1000.
Diluted antibody can be frozen and reused several times.
In IF, the antibody gives unspecific staining of the nucleus. Not cool!
On October 29, 2017 Heidi Gytz Olesen, McGill University, Canada wrote:
THE His Tag Antibody, Genscript A00186-SZ
Short protocol:
1:5000 dilution in 5% low fat milk powder in PBS-T and incubate overnight at 4°C.
Dynabeads protein A kits (Invitrogen) were used for IP. Longer incubation of the antibody in the Ab binding and washing buffer of 1 h was used for optimal binding.
Review:
Worked perfectly for both immunoblotting and co-IP. The diluted antibody could be stored at -20°C after use and be reused 5-10 times for immunoblotting.
On March 16, 2016 Sune Skeldal, Aarhus University wrote:
Good antibody against His-tag for western blotting. Detected his-tagged constructs expressed in media from CHO cells with good result. I used 1:2500 dilution.
On November 4, 2015 Camilla Gustafsen, Aarhus University wrote:
This antibody (1:2500) specifically recognizes 6His tagged PCSK9 secreted to the media of stably transfected HEK293 cells. No band is detected in the lane loaded with media from cells transfected with un-tagged PCSK9. However, a strong background signal is detected from the SeeBlue Plus2 pre-stained protein marker. See inserted figure
On June 3, 2015 Camilla Gustafsen, Aarhus University wrote:
We have used this antibody for WB detection of His tagged PCSK9 produced in CHO cells.
Great result. No background signal
We dilute 1:2500 in 5 % milk buffer and reuse several times
On February 14, 2012 Simon Glerup, Aarhus wrote:
Reasonable for detecting His-tagged proteins by Western blotting.
WB dilution 1:1000.
Diluted antibody can be frozen and reused several times.
In IF, the antibody gives unspecific staining of the nucleus. Not cool!
Good for IP of His-tagged proteins.